Objectives: The aim of this study was to examine changes in the dentinal fluid flow (DFF) during desensi- tizing agent application and to compare permeability after application among the agents. Materials and Methods: A Class 5 cavity was prepared to exposure cervical dentin on an extracted human premolar which was connected to a sub-nanoliter fluid flow measuring device (NFMD) under 20 cm water pressure. DFF was measured from before application of desensitizing agent (SealP SuperSeal, SS; BisBlock, BB; Gluma desensitizer, GL; Bi-Fluoride 12, BF) through application procedure to 5 min after application. Results: DFF rate after each desensitizing agent application was significantly reduced when compared to initial DFF rate before application ( p < 0.05). SP showed a greater reduction in DFF rate than GL and BF did (p < 0.05). SS and BB showed a greater reduction in DFF rate than BF did (p < 0.05). Conclusions: Characteristic DFF aspect of each desensitizing agent was shown in NFMD during the applica- tion procedure. (J Kor Acad Cons Dent 2010;35(5):313-320.) ABSTRACT